Hybrid corn plant and seed pp59601

ABSTRACT

This invention provides hybrid maize plant designated PP59601. This invention further provides hybrid seed of PP59601, hybrid plants produced from such seed, and variants, mutants, and trivial modifications to hybrid PP59601, as well as methods of using the hybrid and products produced from the hybrid.

FIELD OF THE INVENTION

This invention is in the field of maize breeding, specifically relatingto an amylose hybrid maize designated PP59601.

BACKGROUND OF THE INVENTION

All publications and patent applications herein are incorporated byreference for all purposes to the same extent as if each individualpublication or patent application was specifically and individuallyindicated to be incorporated by reference.

The following description includes information that may be useful inunderstanding the present invention. It is not an admission that any ofthe information provided herein is prior art or relevant to thepresently claimed inventions, or that any publication specifically orimplicitly referenced is prior art.

The goal of plant breeding is to combine in a single variety or hybridvarious desirable traits, or to provide a desirable trait withoutsignificant detriment to other important properties. For field crops,desirable traits may include resistance to diseases and insects,tolerance to heat, cold and drought, reducing the time to crop maturity,greater yield, and better agronomic quality. With mechanical harvestingof many crops, uniformity of plant characteristics such as germinationand stand establishment, growth rate, maturity, and plant and ear heightis important. Other desirable traits may be those directly or indirectlyassociated with special nutritional and industrial types of crops.Examples of such specialty varieties or hybrids include those withhigher oil content, different oil profiles, greater protein content,better protein quality or higher amylose content. It is also desirableto produce plants which are particularly adapted to a given agriculturalregion. New hybrids are an important part of efforts to control rawmaterial costs.

Maize (Zea mays L.) is often referred to as corn in the United States,and the terms are used interchangeably in the present application. Maizehas separate male and female flowers on the same plant, located on thetassel and the ear, respectively. Thus, it can be bred by crossing toitself (self-pollination or selfing), to another plant of the samefamily, line or variety (sib-pollination or sib-crossing) or to anotherplant of a different family, line or variety (outcrossing orcross-pollination).

Repeated self-pollination of plants, combined with selection for thedesired type over many generations, results in inbred lines which arehomozygous at almost all loci and thus will produce a uniform populationof homozygous offspring when subject to further self-pollination. Across between two different homozygous lines produces a uniformpopulation of heterozygous hybrid plants. A cross of two plants eachheterozygous at a number of gene loci will produce a population ofheterogeneous plants that differ genetically and will not be uniform.

Hybrid maize varieties can be produced by a process comprising (1) theselection of plants from various germplasm pools for initial breedingcrosses; (2) the selfing of the selected plants from the breedingcrosses for several generations to produce a series of inbred lines asdescribed above; and (3) crossing a selected inbred line with adifferent inbred line to produce the hybrid progeny (F1). Preferably, aninbred line should comprise homozygous alleles at about 95% or more ofits loci.

Pedigree breeding and recurrent selection are two examples of methodsused to develop an inbred line.

Pedigree breeding starts with the crossing of two or more genotypes,each of which may have one or more desirable characteristics. Superiorprogeny are selfed and selected in successive generations, during thecourse of which the level of homozygosity is increased. An inbred linesuitable for hybrid production may be produced after a number ofgenerations of selfing and selection, for example after four, five, sixor more generations.

Double haploid methods can reduce the number of generations needed toobtain an inbred line. These methods involve the doubling of haploidsderived from either the maternal or paternal gametes. Genetics markerscan be used to identify haploids, and the haploids doubled to formhomozygous diploid lines.

Recurrent selection entails individual plants cross-pollinating witheach other to form progeny which are then grown. The superior progenyare then selected by any number of methods, which include individualplant, half sib progeny, full sib progeny, selfed progeny andtopcrossing. The selected progeny are cross pollinated with each otherto form progeny for another population. This population is planted andagain superior plants are selected to cross pollinate with each other.The objective of this repeated process is to improve the traits of apopulation. The improved population can then be used as a source ofbreeding material to obtain inbred lines to be used in hybrids.

Backcrossing can be used to improve inbred lines and a hybrid which ismade using those inbreds. Backcrossing can be used to transfer aspecific desirable trait from one line, the donor parent, to an inbredcalled the recurrent parent which has overall good agronomiccharacteristics yet that lacks the desirable trait. This transfer can beachieved by first crossing the recurrent parent with the donor parent,and then performing a backcross in which the progeny are mated to therecurrent parent. The resultant progeny can then be selected for thedesired trait, and a further backcross performed using the selectedindividuals. Typically after four or more backcross generations withselection for the desired trait in each generation, the progeny willcontain essentially all genes of the recurrent parent except for thegenes controlling the desired trait. The last backcross generation isthen selfed to give pure breeding progeny for the gene(s) beingtransferred.

Other plant breeding techniques known in the art, such as restrictionfragment length polymorphism enhanced selection, genetic marker enhancedselection and transformation, may also be used in the production ofinbred lines. For example, selection in the breeding process can bebased upon the accumulation of markers linked to the positive effectingalleles and/or the elimination of markers linked to the negativeeffecting alleles from the plant's genome. Often, a combination oftechniques is used.

For a review of plant breeding methods well known to those skilled inthe art, see, for example, Sprague and Dudley (eds.), Corn and CornImprovement, Third Edition, American Society of Agronomy, Inc., 986pages, 1988; Fehr and Hadley (eds.), Hybridization of Crop Plants,American Society of Agronomy, Inc., 765 pages, 1980; Allard, Principlesof Plant Breeding, John Wiley & Sons, Inc., 485 pages, 1960; Jensen,Plant Breeding Methodology, John Wiley & Sons, Inc., 676 pages, 1988;Simmonds, Principles of Plant Breeding, Longman Group Limited, 408pages, 1979; and Hallauer and Miranda, Quantitative Genetics in MaizeBreeding, Iowa State University Press, 468 pages, 1981.

In producing a hybrid strain by crossing two different inbred lines, itis advantageous to minimize the possibility of self-pollination.Minimizing self-pollination will minimize the proportion of theresultant seed which is substantially identical to the inbred line(resulting from the self-pollination) and increase the amount of hybridseed (resulting from cross pollination). To this end, commercial maizehybrid production uses a male sterility system to render the femaleparent male sterile. There are several ways in which a maize plant canbe manipulated so that it is male sterile. These include use of manualor mechanical emasculation (or detasseling), cytoplasmic genetic malesterility, nuclear genetic male sterility or gametocides (chemicalagents affecting cells critical to male fertility, for example asdescribed in Carlson, Glenn R., U.S. Pat. No. 4,936,904).

In detasseling, alternate strips of two inbred varieties of maize areplanted in a field, and the pollen-bearing tassels are removed from oneof the inbreds (female) prior to pollen shed. Providing that there issufficient isolation from sources of foreign maize pollen, the ears ofthe detasseled inbred will be fertilized only from the other inbred(male), and the resulting seed is therefore hybrid and will form hybridplants.

Alternatively, the female line can be cytoplasmic male sterile as aresult of an inherited factor in the cytoplasmic genome. Thischaracteristic is inherited exclusively through the female parent inmaize plants, since only the female provides cytoplasm to the fertilizedseed. CMS plants are fertilized with pollen from another inbred that isnot male-sterile. Pollen from the second inbred may or may notcontribute genes that make the hybrid plants male-fertile. The samehybrid seed, a portion produced from detasseled fertile maize and aportion produced using the CMS system can be blended to insure thatadequate pollen loads are available for fertilization when the hybridplants are grown.

Genetic male sterility may be conferred by one of several availablemethods, such as multiple mutant genes at separate locations within thegenome that confer male sterility, as disclosed in U.S. Pat. Nos.4,654,465 and 4,727,219 to Brar et al. and chromosomal translocations asdescribed by Patterson in U.S. Pat. Nos. 3,861,709 and 3,710,511. Asystem in which male fertility genes are expressed under an induciblepromoter is described in Albertsen et al., U.S. Pat. No. 5,432,068.Other approaches include delivering into the plant a gene encoding acytotoxic substance associated with a male tissue specific promoter, oran antisense system in which a gene critical to fertility is identifiedand an antisense to that gene is inserted in the plant (see Fabinjanski,et al. EPO 89/3010153.8 publication no. 329,308 and PCT applicationPCT/CA90/00037 published as WO 90/08828).

Having obtained a desirable hybrid strain by the crossing of twodifferent parent inbred strains, it is possible to maintain a uniformsupply of the hybrid seed. The population of parent plants can bemaintained by repeated self pollination. Moreover, since the parents arehomozygous, the hybrid produced in the cross will always be the same.Thus, once a desirable hybrid has been identified, a continual supply ofhybrid seed having the same properties can be provided.

Objectives of commercial maize hybrid line development include thedevelopment of new corn hybrids which are able to produce high yield ofgrain, which require less investment of time or resources, which aremore resistant to environmental stresses (e.g., stresses particular to acertain growing area), which are easier to harvest and/or which providegrain or other products particularly suitable for a desired commercialpurpose. To obtain a new hybrid, the corn breeder selects and developssuperior inbred parental lines for producing hybrids. This is far fromstraightforward in view of the number of segregating genes and in viewof the fact that the breeder often does not know the desired parentalgenotype in detail. Then, the breeder must identify the particularcross-combination of inbred lines which produces a desired hybrid. Evenhaving obtained two superior inbred lines, there is no guarantee thatthe combination of these will produce desirable hybrid F1 plants. Thisis particularly the case because many selectable traits (e.g., yield)are dependent on the effects of numerous genes interacting with eachother. Thus, the selection or combination of two parent lines produces aunique hybrid which differs from that obtained when either of theparents is crossed with a different inbred parent line.

SUMMARY OF THE INVENTION

This invention relates to the development of a new amylose maize hybriddesignated as PP59601. PP59601 is higher yielding than currently-grownamylose maize hybrids of similar maturity, type and adaptation. Forexample, PP59601 yielded 8.1 bushels more per acre than the mean yieldof a current commercial hybrid when tested together at 4 locations inone year. The yield differential was 3 bushels per acre in trials at 4locations the following year. PP59601 has comparable harvest moisture toother commercial amylose hybrids of similar maturity, improved stalkquality and is a soft grain type suitable for wet milling. PP59601further provides corn growers with a new amylose maize hybrid with highagronomic yield that is adapted to the central corn growing belt of theUnited States.

According to the invention, there is provided a novel corn hybrid,designated PP59601, produced by crossing vA207 and vBL258. These twoproprietary inbreds were developed by the pedigree breeding method.Inbreds vA207 and vBL258 are respectively the female and male parents ofhybrid PP59601. A representative sample of seed which when grownproduces hybrid plants of PP59601 is deposited under American TypeCulture Collection (“ATCC”) accession number PTA-9639.

In one aspect, the present invention provides hybrid seed, arepresentative sample of which has been deposited under ATCC accessionnumber PTA-9639. The present invention also provides a population ofcorn seeds, wherein at least 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%,88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% ofsaid seeds are hybrid seeds of which a representative sample has beendeposited under ATCC accession number PTA-9639.

In another aspect, the present invention relates to a hybrid plantobtainable or obtained by growing seed of which a representative sampleis deposited under ATCC accession number PTA-9639.

The invention also relates to variants, mutants and trivialmodifications of the hybrid seed or plant.

Seeds, plants, plant parts, somatic tissues or cells according to thepresent invention may have substantially the same genotype as thedeposited seed ATCC PTA-9639, and/or may be capable of serving as thesource for tissue culture to produce a plant of substantially the samegenotype as hybrid seed deposited under ATCC accession number PTA-9639.

In another aspect the present invention provides a corn plant (or seedthereof) having desirable traits of hybrid PP59601. The corn plant mayhave all or essentially all of the morphological or physiologicalcharacteristics of hybrid PP59601. Optionally, the plant may have one ormore additional characteristics, e.g., characteristics resulting fromthe presence of one or more nucleic acid sequences introduced bytechniques known to those skilled in the art, such as transgenictechniques or conventional breeding methods such as backcrossing. Inother words, the hybrid corn plants of the present invention includehybrid corn plants of PP59601 which further include one, two, three ormore foreign or heterologous genes introduced into PP59601. Such foreignor heterologous genes may be from a different corn plant (i.e., a corninbred, corn hybrid, corn haploid, etc.) other than the inbreds used toproduce PP59601, and/or from a plant species other than Zea mays (e.g.,alfalfa, soybean, canola, tomato, potato, yew tree, marigold, etc.),and/or from a non-plant species (e.g., bacteria, fungi, insects,mammals, jellyfish, etc.).

The invention further relates to corn plants and seeds derived fromhybrid maize PP59601. These plants and seeds may be of an essentiallyderived variety as defined in section 41(3) of the Plant VarietyProtection Act, i.e., a variety that:

-   -   (i) is predominantly derived from hybrid PP59601 or from a        variety that is predominantly derived from hybrid PP59601, while        retaining the expression of the essential characteristics that        result from the genotype or combination of genotypes of hybrid        PP59601;    -   (ii) is clearly distinguishable from hybrid PP59601; and    -   (iii) except for differences that result from the act of        derivation, conforms to the initial variety in the expression of        the essential characteristics that result from the genotype or        combination of genotypes of the initial variety.

An essentially derived variety may be obtained by the selection of anatural or induced mutant or of a somaclonal variant, the selection of avariant individual from plants of hybrid PP59601, backcrossing,transformation by genetic engineering, or any other method.

The essential characteristics may be one or more of the desirable traitsset forth herein.

The corn plants and seeds derived from hybrid maize PP59601 may in otherembodiments be regenerated from a tissue culture produced from a hybridPP59601 plant, or may be a plant or seed having hybrid PP59601 as anancestor, as discussed further below.

The present invention also provides a tissue culture of regeneratablecells produced from hybrid plant PP59601, wherein said tissue culture iscapable of producing plants having desirable traits of hybrid PP59601 asset out above. The tissue culture may be derived directly or indirectlyfrom hybrid PP59601. Preferably the tissue culture is capable ofproducing plants which have all or substantially all of themorphological and physiological characteristics of hybrid PP59601.Optionally, the plants may have one or more additional characteristic,e.g., conferred by a nucleic acid sequence introduced using transgenicor conventional breeding techniques. In some embodiments the plant mayhave the genetic complement of hybrid PP59601, optionally comprising oneor more additional nucleic acid sequences capable of modifying thephenotype of the plant when expressed (e.g., as RNA or protein). Theculture can be from any tissue capable of somatic embyrogenesis, e.g.,may be selected from the group consisting of leaf, pollen, embryo, root,root tip, anther, silk, flower, kernel, ear, cob, husk, stalk, cell orprotoplast.

The invention further relates to the use of the tissue culture toproduce a whole plant, to protoplasts produced from said tissue cultureand to a corn plant regenerated from said tissue culture. A method ofproducing a whole plant from the tissue culture may comprise one or moreof: culturing cells in vitro in a media comprising an embryogenesispromoting hormone until callus organization is observed; transferringcells to a media which includes a tissue organization promoting hormone;after tissue organization is observed transferring cells into a mediawithout said hormone to produce plantlets; and growing said plantlets,optionally including growing said plantlets on a minimal media forhardening.

In a further aspect of the present invention, there is provided pollenor an ovule of hybrid plant PP59601, as well as seed produced byfertilization with said pollen or of said ovule, and plants grown fromthe seed.

The hybrid plant PP59601 can be crossed with a corn plant of anotherline or variety, or can be sib-crossed or selfed to produce anotherplant, line (e.g., inbred line) or population of plants (e.g., breedingpopulation of plants) which is of benefit in plant breeding.

Thus, in another aspect the present invention relates to a plant or seedproduced by a breeding program using hybrid PP59601 as a parent, whereinthe plant or seed is a member of a generation of progeny of said parent,e.g., a member of the first, second, third, fourth, fifth, sixth or moregeneration of progeny. Thus, the present invention includes plants andseeds produced using hybrid PP59601 as an ancestor. Ancestry can beassessed from the records kept routinely by one of ordinary skill in theart. It can also be assessed based on nucleic acid identity, e.g., usingmolecular markers, electrophoresis and the like. The plant or seed thusproduced may have desired characteristics of hybrid PP59601 as discussedabove, or may have all of the morphological and physiological traits ofhybrid PP59601.

In another aspect the present invention relates to use of a hybridPP59601 maize plant to produce seed and/or progeny maize plants. Thepresent invention also provides a method comprising providing a plant ofhybrid PP59601, crossing it with itself or with another maize plant(which may be another hybrid PP59601 plant or may be a plant of adifferent line or variety) so as to produce seed, and harvesting saidseed. The method may further comprise growing said seed to produce oneor more progeny maize plants, and optionally, breeding from one or moreof said progeny maize plants to produce progeny seed, which may beharvested. The step of growing the progeny seed and breeding from theresultant maize plants to produce a further population of seed can berepeated over one or more further generations (e.g., in 1, 2, 3, 4, 5, 6or more further generations). For instance, the progeny may be selfed,sibbed, backcrossed, crossed to a population or the like. By “breedingfrom” a plant is meant a process of crossing the plant with itself orwith another plant of the same or a different variety to produce seed.Selection may be carried out in one or more of the progeny generations.The selection may be for one or more desirable traits of hybrid PP59601,e.g., one or more of amylose content of the starch and agronomic yield.Selection may be done using visual inspection, or using molecularmarkers.

Plants resulting from such methods would contain desirable traitsderived from hybrid PP59601 and thus would benefit from the work of thepresent inventors and from the disclosure contained herein.

For instance, in one embodiment, a method of the invention may comprisesib or self-pollinating hybrid PP59601 to produce a first generation ofprogeny plants. The method may further comprise sib or self-crossingsaid progeny over one or more further generations (e.g., 1, 2, 3, 4, 5,6 or more further generations) and/or double haploid breeding, in orderto produce a plant which is substantially homozygous, e.g., greater than90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.1%, 99.2%, 99.3%,99.4%, 99.5%, 99.6%, 96.7%, 99.8%, 99.9%, 99.95% or more homozygous.This method may comprise selection of plants having the one or moredesirable traits of the parent plant. This selection may take place ineach progeny generation or less frequently, e.g., in 1, 2, 3, 4, 5 ormore generations of progeny (e.g., in the first progeny generationand/or in one or further progeny generations.

In another embodiment, a hybrid maize plant as described herein can alsobe crossed to a different variety of maize, such as an inbred line(e.g., an elite inbred line). The F1 progeny generation resulting fromthis cross would have 50% of its genes derived from the hybrid PP59601.The method may further comprise self-fertilization of one or more plantsfrom the F1 population to produce an F2 progeny generation. Some of theF2 plants will by chance have more than 50% of their genes derived fromthe parental hybrid plant. These may be selected, for example usingmolecular marker selection or selection of one or more desired traits ofhybrid PP59601. Self-fertilization of the progeny may be carried outover 1, 2, 3, 4, 5 or more further generations to produce an inbredline. Selection may be carried out in each progeny generation, or at alower frequency, e.g., in 1, 2, 3, 4, 5 or more of the generations.

The method may in some embodiments further comprise modification of theresultant inbred line to provide a further desired trait or traits. Forinstance, the method may comprise crossing the resultant inbred linewith a further plant variety having a desirable trait, and backcrossingthe progeny over 1, 2, 3, 4, 5, 6 or more generations so as to insertthe desired trait into a genetic background which is substantially thatof the inbred line. In another embodiment, the method may comprisetransgenic modification of the inbred line, which can be carried outusing methods which would be well known to those in the art.

In a further embodiment the method comprises crossing a plant of a firstvariety or line to a plant of a second, different variety or line,wherein the first variety or line is hybrid PP59601. The second varietyor line may be an inbred line and in some embodiments, may be of one ofthe parental lines of hybrid PP59601. The method may comprise growing afirst progeny generation. The method may then further comprisebackcrossing one or more plants of that progeny generation to one ormore plants of the second variety or line to produce a further progenygeneration. The backcrossing may be repeated in 1, 2, 3, 4, 5, 6 or moregenerations. The last backcross generation may be selfed to result in apure breeding line for the desired trait(s). Selection may be carriedout in one or more of the progeny populations, e.g., to select plantshaving one or more desirable traits of hybrid PP59601.

The invention also includes the population of seeds or plants producedat any stage of the breeding methods described above. In someembodiments, the seed or plant may be an inbred seed or plant, e.g.,such as may be used for a further breeding program or for thedevelopment of further hybrids.

Corn is a highly useful crop, and numerous commercial products can beprovided by or derived from its different parts. Accordingly, thepresent invention provides use of a plant as described herein for theproduction of a processed corn product.

Also provided is a method comprising providing one or more parts of aplant as described herein and processing said part(s) to produce aprocessed corn product. The method may also comprise growing the plantand/or harvesting said one or more parts.

The plant part may be any of the parts described above, including thestem, husk or cob, but in many embodiments will be the ear or thekernels.

Examples of processed corn products are corn starch (including isolatedcorn starch components such as amylose or amylopectin), flour, grits,meal, corn syrup or dextrose, corn oil, processed corn grain productssuch as canned, frozen or pureed grain, ethanol, paper, wall-board orcharcoal.

For instance, in one embodiment the invention provides a method for theproduction of corn starch comprising providing kernels of a plant asdescribed herein, and processing the kernels to produce corn starch. Theprocessing may comprise wet-milling.

In another embodiment, the invention provides a method for theproduction of corn flour comprising providing kernels of a plant asdescribed herein, and processing the kernels to produce corn flour. Theprocessing may comprise dry-milling.

The invention also provides a method comprising, having provided aprocessed corn product as described above, using said processed cornproduct in the production of a manufactured product. These may be any ofthe manufactured products as described further below. Examples include afood product, packaging, adhesive, paper or textile, pharmaceuticalproduct, cosmetic, and home care product.

The invention further provides a processed corn product or manufacturedproduct produced by any of the methods described above. A preferredprocessed corn product may be high amylose starch or flour.

Definitions

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can be used inthe practice or testing of the present invention, the preferred methodsand materials are described.

As used herein, the term “allele” refers to any of several alternativeforms of a gene.

As used herein, “starch” refers to starch in its natural or native formas well as also referring to starch modified by physical, chemical,enzymatic and biological processes.

As used herein, “amylose” refers to a starch polymer that is anessentially linear assemblage of D-anhydroglucose units which are linkedby alpha 1,6-D-glucosidic bonds.

As used herein, “amylose content” refers to the percentage of theamylose type polymer in relation to other starch polymers such asamylopectin.

As used herein, “area of adaptation” refers to an area having aparticular combination of environmental conditions under which this cornhybrid will grow well. The term is not intended to mean that the cornhybrid will not grow outside of this region, particularly, that it willnot grow equally well in areas sharing the same or substantially thesame combination of conditions.

As used herein, “high amylose maize” or “amylomaize” refer to thegeneric name for corn that has an amylose content of about 50% orgreater. The single recessive amylose-extender gene (ae1), plusmodifiers, gives a range in amylose content of about 50% to about 94%.Amylomaize hybrids require special management and cultural requirementsto provide more assurance of optimum grain production of acceptablequality and purity. Production fields must be isolated from normal dentcorn. High-amylose grain is grown exclusively under contract for wet anddry milling. Amylose starch is utilized in a complexity of uses invarious industries. Similar to yield, the actual amylose content of asample of grain from a particular variety in any particular trial canvary slightly from its overall mean or median amylose content dependingon the particular environment in which it is grown. As known by thoseskilled in the art of growing maize, many factors are involved indetermining what constitutes a particular environment for a particulartrial/growing season (e.g., rainfall, temperature, soil type, diseaseincidence, cloud cover, etc.).

As used herein, “amylose maize inbred” refers to maize inbred that hasan amylose content of about 50% or greater, wherein the amyloseconcentration of the grain is determined by the calorimetric method.

As used herein, “amylose maize hybrid” refers to maize hybrid that hasan amylose content of about 50% or greater, wherein the amyloseconcentration of the grain is determined by the calorimetric method.

As used herein, the terms “crossing ” or “crossed” or grammaticalequivalents thereof refer to pollen from one flower being transfers tothe ovule of the same or a different flower to result in fertilization.A plant crossed to itself is self-pollinated or selfed; a plant crossedto another plant of the same variety, family or line is sib-pollinatedor sib-crossed and a plant crossed to another plant of a differentvariety, family or line is out-crossed or out-pollinated.

As used herein, the term “cross pollination” or “cross-breeding” referto the process by which the pollen of one flower on one plant is applied(artificially or naturally) to the ovule (stigma) of a flower on anotherplant.

As used herein, the term “cultivar” refers to a variety, strain or raceof plant that has been produced by horticultural or agronomic techniquesand is not normally found in wild populations.

As used herein, the term “elite inbred line” refers to an inbred whichhas been shown to contribute desirable qualities when used to producecommercial hybrids.

As used herein, the term “female” refers to a plant that producesovules. Female plants generally produce seeds after fertilization. Aplant designated as a “female plant” may contain both male and femalesexual organs. Alternatively, the “female plant” may only contain femalesexual organs either naturally (e.g., in dioecious species) or due toemasculation (e.g., by detasselling).

As used herein, the term “filial generation” refers to any of thegenerations of cells, tissues or organisms following a particularparental generation. The generation resulting from a mating of theparents is the first filial generation (designated as “F1” or “F₁”),while that resulting from crossing of F1 individuals is the secondfilial generation (designated as “F2” or “F₂”).

As used herein, the term “gamete” refers to a reproductive cell whosenucleus (and often cytoplasm) fuses with that of another gamete ofsimilar origin but of opposite sex to form a zygote, which has thepotential to develop into a new individual. Gametes are haploid and aredifferentiated into male and female.

As used herein, the term “gene” refers to any segment of DNA associatedwith a biological function. Thus, genes include, but are not limited to,coding sequences and/or the regulatory sequences required for theirexpression. Genes can also include nonexpressed DNA segments that, forexample, form recognition sequences for other proteins. Genes can beobtained from a variety of sources, including cloning from a source ofinterest or synthesizing from known or predicted sequence information,and may include sequences designed to have desired parameters. Thus,this invention further encompasses the maize plants, and parts thereof,of the present invention which have been transformed so that its geneticmaterial contains one or more transgenes operably linked to one or moreregulatory elements. Furthermore, the maize plants, or parts thereof, ofthe present invention also encompass such maize plants, or partsthereof, that contain a single gene conversion.

As used herein, the term “genetic complement” refers to the complete setof alleles possessed by a cell. In a plant or other somatic tissue orcell the complement will be diploid—that is, there will be two alleles(the same or different) at each locus.

As used herein, the term “genotype” refers to the genetic makeup of anindividual cell, cell culture, tissue, plant, or group of plants.

As used herein, the term “grain” refers to mature corn kernels producedby commercial growers for purposes other than growing or reproducing thespecies.

As used herein, the terms “heterologous polynucleotide” or a“heterologous nucleic acid” or an “exogenous DNA segment” refer to apolynucleotide, nucleic acid or DNA segment that originates from asource foreign to the particular host cell, or, if from the same source,is modified from its original form. Thus, a heterologous gene in a hostcell includes a gene that is endogenous to the particular host cell, buthas been modified. Thus, the terms refer to a DNA segment which isforeign or heterologous to the cell, or homologous to the cell but in aposition within the host cell nucleic acid in which the element is notordinarily found. Exogenous DNA segments are expressed to yieldexogenous polypeptides.

As used herein, the term “heterologous trait” refers to a phenotypeimparted to a transformed host cell or transgenic organism by anexogenous DNA segment, heterologous polynucleotide or heterologousnucleic acid.

As used herein, the term “heterozygote” refers to a diploid or polyploidindividual cell or plant having different alleles (forms of a givengene) present at least at one locus.

As used herein, the term “heterozygous” refers to the presence ofdifferent alleles (forms of a given gene) at a particular gene locus.

As used herein, the term “homologue” refers to a nucleic acid or peptidesequence which has a common origin and functions similarly to a nucleicacid or peptide sequence from another species.

As used herein, the term “homozygote” refers to an individual cell orplant having the same alleles at one or more loci.

As used herein, the term “homozygous” refers to the presence ofidentical alleles at one or more loci in homologous chromosomalsegments.

As used herein, the term “hybrid” refers to any individual cell, tissueor plant resulting from a cross between parents that differ in one ormore genes.

As used herein, the term “inbred” or “inbred line” refers to arelatively true-breeding strain.

As used herein, the term “kernel” refers to the corn caryopsiscomprising a mature embryo and endosperm which are products of doublefertilization.

As used herein, the term “line” is used broadly to include, but is notlimited to, a group of plants vegetatively propagated from a singleparent plant, via tissue culture techniques or a group of inbred plantswhich are genetically very similar due to descent from a commonparent(s). A plant is said to “belong” to a particular line if it (a) isa primary transformant (T0) plant regenerated from material of thatline; (b) has a pedigree comprised of a T0 plant of that line; or (c) isgenetically very similar due to common ancestry (e.g., via inbreeding orselfing). In this context, the term “pedigree” denotes the lineage of aplant, e.g. in terms of the sexual crosses effected such that a gene ora combination of genes, in heterozygous (hemizygous) or homozygouscondition, imparts a desired trait to the plant.

As used herein, the term “locus” (plural: “loci”) refers to any sitethat has been defined genetically. A locus may be a gene, or part of agene, or a DNA sequence that has some regulatory role, and may beoccupied by the same or different sequences.

As used herein, the term “male” refers to a plant that produces pollengrains. The “male plant” generally refers to the sex that producesgametes for fertilizing ova. A plant designated as a “male plant” maycontain both male and female sexual organs. Alternatively, the “maleplant” may only contain male sexual organs either naturally (e.g., indioecious species) or due to removal of the ovary.

As used herein, the term “mass selection” refers to a form of selectionin which individual plants are selected and the next generationpropagated from the aggregate of their seeds.

As used herein, the term “open pollination” refers to a plant populationthat is freely exposed to some gene flow, as opposed to a closed one inwhich there is an effective barrier to gene flow.

As used herein, the terms “open-pollinated population” or“open-pollinated variety” refer to plants normally capable of at leastsome cross-fertilization, selected to a standard, that may showvariation but that also have one or more genotypic or phenotypiccharacteristics by which the population or the variety can bedifferentiated from others. A hybrid, which has no barriers tocross-pollination, is an open-pollinated population or anopen-pollinated variety.

As used herein, the term “ovule” refers to the female gametophyte,whereas the term “pollen” means the male gametophyte.

As used herein, the term “phenotype” refers to the observable charactersof an individual cell, cell culture, plant, or group of plants whichresults from the interaction between that individual's genetic makeup(i.e., genotype) and the environment.

As used herein, the term “recombinant” or “recombinants” refer to acell, tissue or organism that has undergone transformation withrecombinant DNA. The original recombinant is designated as “R0” or “R₀.”Selfing the R₀ produces a first transformed generation designated as“R1” or “R₁.”

The term “plants” or “plant” or grammatical equivalents thereof as usedherein is to be construed broadly to include, as well as whole organisms(i.e., plants, also sometimes called whole plants) at any stage of theirdevelopment, plant cells, plant protoplasts, tissue culture, plantcalli, plant embryos or parts of a plant such as roots, root tips,stalk, leaves, flowers, anthers, ears, cobs, husks, silks, and kernels.

As used herein, the term “seed” refers to mature corn kernels producedfor the purpose of propagating the species.

As used herein, the term “self pollinated” or “self-pollination” meansthe pollen of one flower on one plant is applied (artificially ornaturally) to the ovule (stigma) of the same or a different flower onthe same plant.

As used herein, “MST PCT” refers to the actual moisture of grain atharvest.

As used herein, “PER CENT DROPPED EARS” refers to the percentage of earsof corn that have detached from the plant and fallen to the ground.

As used herein, “PLTPOP” refers to the percentage of plants which haveemerged after planting in comparison to the mean percentage of allhybrids in a common test.

As used herein, “staygreen” refers to a measure of plant health that isdetermined by the percentage of green tissue compared to desiccatedbrown tissue on the plant at physiological maturity.

As used herein, “drydown” or “dry down” refer to loss of grain moistureover time.

As used herein, “STKLOD PCT” refers to the percentage of plants in whichthe stalk is broken below the ear node.

As used herein, “TST/WT LB/BU” refers to a measure of the grain weightin pounds for a given bushel volume.

As used herein, the term “synthetic” refers to a set of progeniesderived by intercrossing a specific set of clones or seed-propagatedlines. A synthetic may contain mixtures of seed resulting from cross-,self-, and/or sib-fertilization.

As used herein, the term “transformation” refers to the transfer ofnucleic acid (i.e., a nucleotide polymer) into a cell. As used herein,the term “genetic transformation” refers to the transfer andincorporation of DNA, especially recombinant DNA, into a cell.

As used herein, the term “transformant” refers to a cell, tissue ororganism that has undergone transformation. The original transformant isdesignated as “T0” or “T₀.” Selfing the T0 produces a first transformedgeneration designated as “T1” or “T₁.”

As used herein, the term “transgenic” refers to cells, cell cultures,organisms, plants, and progeny of plants which have received a foreignor modified gene by one of the various methods of transformation,wherein the foreign or modified gene is from the same or differentspecies than the species of the plant, or organism, receiving theforeign or modified gene.

As used herein, the term “variety” refers to a subdivision of a species,consisting of a group of individuals within the species that aredistinct in form or function from other similar arrays of individuals.

DETAILED DESCRIPTION OF THE INVENTION

PP59601 is a cross between the female inbred vA207 by the male inbredvBL258. Inbreds vA207 and vBL258 were developed using the pedigreemethod of plant breeding. VA207 is a B73 derived inbred while vBL258 isderived from an LH123 background. Both vA207 and vBL258 have amyloseconcentration between 55 and 58%.

Hybrid PP59601 is characterized by high agronomic yield, averagedrydown, and good stalk quality. Amylose maize hybrids are generallyagronomically inferior to other types of maize hybrids such as dent andwaxy grown for commercial production. Hybrid PP59601 has a relativematurity of approximately 112 days based on the comparative relativematurity system for grain harvest moisture. It is adapted to the eastcentral corn belt regions of Indiana and Ohio. The hybrid has thefollowing characteristics based on data collected from field plotslocated in Lebanon, Ind.

TABLE 1 Variety Description Information for PP59601 A. Type: 2 (1 =Sweet 2 = dent 3 = Flint 4 = Flour 5 = Pop) Pedigree: vA207 × vBL258 B.Maturity: Days Heat Units 74 1510 From plant emergence to 50% of plantswith pollen 75 1525 From Plant emergence to 50% of plants with silk C.Plant Characteristics: Standard Sample Deviation Size 227.1 cm PlantHeight (tassel tip) 5.76 10 97.9 cm Ear Height (base of top ear node)6.98 10 0 Average number of tillers/plant 0 10 1.2 Average number ofears/stalk 0.42 10 Root Color Munsell code: 5 RP 6/4 1 Anthocyanin ofbrace roots (1 = absent; 2 = faint; 3 = moderate; 4 = dark; 5 = verydark) D. Leaf Standard Sample Deviation Size 10.6 cm Width of ear nodeleaf 0.52 10 88.9 cm Length of ear node leaf 2.18 10 Leaf Color Munsellcode: 5 G 3/4 open Leaf Arch E. Tassel Standard Sample Deviation Size4.9 Number of primary lateral branches 0.7 10 42.1 cm Tassel length (topleaf collar to tassel tip) 2.47 10 8 Pollen shed (1 = light to 9 =heavy) 0 10 Anther color yellow Munsell code: 5 YR 7/4 Glume colorMunsell code: 5 R 7/6 10.2 cm Peduncle length (top leaf to basalbranches) 0.92 10 upright Tassel Arch F. Ear (unhusked data) Silk color(3 days after emergence) Munsell code: 2.5 GY 8/12 Husk cover (25 daysafter 50% silking) Munsell code: 5 GY 6/6 Dry husk cover (65 days after50% shedding) Munsell code: 5 Y 8/2 horizontal Position of ear at dryhusk stage medium Husk tightness 2.03 cm Husk extension (1 = short(exposed); 2 = medium (<8 cm); 3 = long (8-10 cm beyond ear tip); 4 =very long (>10 cm)) G. Ear (husked data) Standard Sample Deviation Size18.74 cm Ear length 1.38 10 4.78 cm Ear diameter at midpoint 0.21 10220.47 gm Ear weight 22.47 10 17.8 Number of kernel rows 1.14 10 2Kernel rows (1 = indistinct; 2 = distinct) 2 Row alignment (1 =straight; 2 = slightly curved; 3 = spiral) 2 Ear taper (1 = slight; 2 =average; 3 = extreme) H. Kernel (dried) Standard Sample Deviation Size1.35 cm Kernel length 0.06 10 0.76 cm Kernel width 0.06 10 0.36 cmKernel thickness 0.04 10 28 % Round kernels 4 10 1 Aleurone colorpattern (1 = homozygous; 2 = segregating) Aleurone color Munsell code:2.5 Y 8/10 Hard endosperm color Munsell code: 2.5 Y 8/2 high amyloseEndosperm type 25.87 gm Weight per 100 kernels 1.0 10 I. Cob StandardSample Deviation Size 2.95 cm Cob diameter at mid-point 0.41 10 Cobcolor Pink Munsell code: 10 R 5/6 J. Disease resistance (Rate from 1 =most susceptible to 9 = most resistant) 7 Common rust (Puccinia sorghi)6 Grey leaf spot (Cercospora zeae-maydis) 6 Northern leaf blight(Exserohilum turcicum) 5 Southern leaf blight (Bipolaris maydis) 6Stewart's wilt (Erwinia stewarti) K. Insect resistance European cornborer (Ostrinia nubalis) 6 First generation 5 Second generation L.Agronomic traits 5 Staygreen (70 days after anthesis, rating scale 1-9,9 = best) 2 Per cent dropped ears (70 days after anthesis)

Variants, mutants and trivial modifications of the hybrid seed or plantPP59601 are within the scope of the present invention. A trivialmodification may be a modification of the genetic code of the hybridplant which results in a plant having the desirable traits of hybridPP59601, as discussed above, and which preferably has all orsubstantially all of the morphological or physiological characteristicsof the hybrid PP59601.

It may be preferred that a seed or plant, e.g., a variant seed or plant,according to the present invention has a genome with at least 90%, 91%,92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% genetic identitywith the genome of hybrid.

A progeny plant of hybrid PP59601 (in any generation) or a plant derivedfrom hybrid PP59601 may preferably have at least 25%, 30%, 35%, 40%,45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%,87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 99.5%,99.9% or 100% genetic identity with hybrid maize plant PP59601

The genotype of a plant and the degree of genetic identity to hybridPP59601 can be assessed using plant breeder records kept routinely byone of ordinary skill in the art. The genotype can additional oralternatively be assessed using molecular marker techniques, e.g, bygenetic marker profiling.

A genetic marker profile can be obtained by techniques such asRestriction Fragment Length Polymorphism (RFLP), Randomly AmplifiedPolymorphic DNA (RAPD), Arbitrarily Primed Polymerase Chain Reaction(AP-PCR), DNA Amplification Fingerprinting (DAF), Sequence CharacterizedAmplified Regions (SCARs), Amplified Fragment Length Polymorphisms(AFLPs), Simple Sequence Repeats (SSRs) which are also referred to asmicrosatellites, and Single Nucleotide Polymorphisms (SNPs). For examplesee Berry, Don et al “Assessing Probability of Ancestry Using SimpleSequence Repeat Profiles: Applications to Maize Hybrids and Inbreds”Genetics 2002, 161: 813-824.

SSRs are frequently used for mapping purposes. This method is based onrepeated sequences which may be repeated a variable number of times atany given locus, thus giving rise to polymorphism, with the potentialfor multiple alleles. Detection of SSR can be achieved by a number ofmethods, including PCR. The PCR detection is done using two primersflanking the region containing the repeats (such primers are publiclyavailable). Following amplification, markers can be scored by gelelectrophoresis of the amplification products. Scoring of the markergenotype is based on the size of the amplified fragment as measured bymolecular weight, rounded to the nearest integer. Relative values shouldremain constant regardless of the specific primer or precise techniqueused.

Thus, references to percentage genetic identity may be references topercentage molecular marker profile identity. The molecular markerprofile may be an SSR profile. The percentages may refer to the geneticcontribution in the molecular marker profile from hybrid PP59601.

It may be preferred that a seed or plant according to the presentinvention has one or more additional desirable traits and/or one or moreinserted nucleic acid sequences conferring a desirable trait whencompared to hybrid PP59601. The nucleic acid sequence may be have beeninserted into the seed or plant or any progenitor thereof by any of themethods known to one skilled in the art, e.g., by transgenic techniquesor by conventional breeding techniques such as backcrossing. Desirabletraits include, but are not limited to, insect, pest or diseaseresistance, resistance to a herbicide, increased drought or coldresistance, male sterility and modification of the properties of thecorn grain (e.g., modified fatty acid metabolism, decreased phytatecontent, modified carbohydrate composition or the like). The source ofthe nucleic acid may be a plant of the same or different species, or maybe any other organism such as an animal (e.g., an insect), prokaryote,fungus, or a virus. The nucleic acid may also be an artificial nucleicacid, i.e., one not appearing in nature.

Specific examples of such genes would be well known to the skilledperson, but some which could be used include a Bacillus thuringiensisprotein, a plant disease resistance gene, a lectin, a vitamin bindingprotein such as avidin, a protease inhibitor or amylase inhibitor, amutant EPSP or aroA gene, an antisense ACP gene or a phytase encodinggene. The nucleic acids may be any genetic material capable of modifyingthe plant's phenotype, e.g., conferring or improving a desirable trait,when expressed in a plant, including antisense nucleic acids, siRNAs andthe like as well as nucleic acid sequences encoding proteins. Thenucleic acid may also be or comprise an enhancer of a promoter. Examplesof suitable nucleic acids can be found in U.S. Pat. No. 6,777,598, thedisclosure of which is incorporated explicitly by reference.

Transgenic methods are well known to those in the art. Both physical andbiological methods for plant transformation are well known in the art(see, for example, Miki et al, “Procedures for Introducing Foreign DNAinto Plants”, in Methods in Plant Molecular Biology and Biotechnology,Glick, B. R. and Thompson, J. E. Eds (CRC Press, Inc, Boca Raton, 1993)pages 67-88). Expression vectors and in vitro culture methods for plantcell and tissue transformation and regeneration of plants are alsoavailable. See for example Gruber et al “Vectors for PlantTransformation”, in Methods in Plant Molecular Biology andBiotechnology, Glick, B. R. and Thompson, J. E. Eds (CRC Press, Inc,Boca Raton, 1993) pages 89-119, and U.S. Pat. No. 6,118,055.

The present invention also relates in some aspects and embodiments totissue cultures, to the use of these cultures and to methods comprisingproducing plants from these cultures.

Duncan, Williams, Zehr, and Widholm, Planta, (1985)165:322-332 reflectsthat 97% of the plants cultured which produced callus were capable ofplant regeneration. Subsequent experiments with both inbreds and hybridsproduced 91% regenerable callus which produced plants. In a furtherstudy in 1988, Songstad, Duncan & Widholm in Plant Cell Reports (1988),7:262-265 reports several media additions which enhance regenerabilityof callus of two inbred lines. Other published reports also indicatedthat “nontraditional” tissues are capable of producing somaticembryogenesis and plant regeneration. K. P. Rao, et al., Maize GeneticsCooperation Newsletter, 60:64-65 (1986), refers to somatic embryogenesisfrom glume callus cultures and B. V. Conger, et al., Plant Cell Reports,6:345-347 (1987) indicates somatic embryogenesis from the tissuecultures of maize leaf segments. Thus, it is clear from the literaturethat the state of the art is such that these methods of obtaining plantsare, and were, “conventional” in the sense that they are routinely usedand have a very high rate of success.

Tissue culture of maize is described in European Patent Application,publication 160,390, incorporated herein by reference. Maize tissueculture procedures are also described in Green and Rhodes, “PlantRegeneration in Tissue Culture of Maize,” Maize for Biological Research(Plant Molecular Biology Association, Charlottesville, Va. 1982, at367-372) and in Duncan, et al., “The Production of Callus Capable ofPlant Regeneration from Immature Embryos of Numerous Zea MaysGenotypes,” 165 Planta 322-332 (1985).

During the production of hybrid seed, effort is made to prevent selfpollination of the inbred parent lines. This can be done by conferringmale sterility on one of the parent lines by techniques which will beapparent to the skilled person, including the techniques discussedabove. However, in the field, complete male sterility of the femaleparent is extremely difficult to achieve and so in packaged hybrid seed,there is potential for the inclusion of a small amount of the selfedfemale parent even when the female seed is or has been treated so as tobe male sterile. Also, because the male parent is grown next to thefemale parent in the field there is the possibility that the male selfedseed could be unintentionally harvested and packaged with the hybridseed.

Therefore, a population of seeds according to the invention may comprisea majority of seeds produced by hybridization of the two parents, andalso comprises levels of seed produced from the selfed parent strains(equivalent to the inbred male and female parent lines) that would beexpected to result from the normal methods of producing the hybrid. Forexample, the seed population may comprise at least 90%, 91%, 92%, 93%,94%, 95%, 96%, 97%, 98%, 99%, 99.5%, 99.9% or 100% of seed produced fromthe hybridization of the two parents. The amount of the female inbredline (i.e., seed produced from the selfed female parent) may be lessthan 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.4%, 0.3%, 0.2%,0.1% or 0.05%. The amount of the male inbred line (i.e., seed producedfrom the selfed male parent) may be less than 5%, 4%, 3%, 2%, 1%, 0.5%,0.4%, 0.3%, 0.2%, 0.1% or 0.05%.

The self-pollinated plants can be identified and distinguished from thehybrid seed because the self-pollinated plants will be geneticallyequivalent to one of the inbred lines used to produce the hybrid. Due tothe level of homozygosity, they will show decreased vigor when comparedto the hybrid. For instance, inbreds are identified by their lessvigorous appearance for vegetative and/or reproductive characteristics,including shorter plant height, small ear size, ear and kernel shape,cob color, or other characteristics.

Identification of these self-pollinated lines can also be accomplishedthrough molecular marker analyses. See, “The Identification of FemaleSelfs in Hybrid Maize: A Comparison Using Electrophoresis andMorphology”, Smith, J. S. C. and Wych, R. D., Seed Science andTechnology 14, pp. 1-8 (1995), the disclosure of which is expresslyincorporated herein by reference. The inbreds can be identified as beinghomozygous at one or more loci. See also, “Identification of AtypicalPlants in Hybrid Maize Seed by Postcontrol and Electrophoresis” Sarca,V. et al., Probleme de Genetica Teoritica si Aplicata Vol. 20 (1) p.29-42.

INDUSTRIAL APPLICABILITY

Corn has extensive use as animal feed, in providing food for humanconsumption, and in providing raw materials for industry.

Corn, including both grain and non-grain portions, is extensively usedas a feed for livestock, such as pigs, cattle and poultry. The grain isalso used for human consumption. In addition, corn kernels can be wetmilled to produce corn starch, corn syrup and dextrose, or can be drymilled to produce corn flour, grits and meal. Corn oil is recovered fromcorn germ, which is a by-product of both the wet and dry millingindustries.

Uses of corn starch are based on functional properties such asviscosity, film formation, adhesive properties and the ability tosuspend particles. Corn starch can be used in industry in the productionof paper, textiles and adhesives. It is also useful in buildingmaterials, foundry binders, laundry starches, explosives, oil-well muds,oil-drilling fluids and other mining applications. Due to theirbiodegradable and renewable nature, starches are increasingly being usedmany other products, including packaging, plastics, detergents,pharmaceutical tablets, pesticides and cosmetics. Starch can also befermented into ethanol and can also be processed into corn syrups andsweeteners such as high fructose corn syrup and dextrose. Starch can beused in an unmodified or modified form (e.g., acid modified corn starch,dextrins, oxidized corn starch, pregelatinized starch and chemicallyderivatized starch).

Corn starch is made up of two components, amylose and amylopectin.Amylose consists of predominantly linear chains of glucose monomerslinked by 1,4-glycosidic bonds. In amylopectin, the chains are branchedby the addition of 1,6-glycosidic bonds. Starches and flours havingdifferent proportions of amylose and amylopectin are particularlyadapted to different industrial purposes.

High amylose starch may be recognized by one or more of the followingproperties. The granules are of two distinct types, spherical andirregular, and are smaller than normal starch granules. TheBirefringence End Point Temperature (“BEPT”) is reported as 97° C. BEPTis the temperature at which the starch molecule loses organizedstructure. Some of the granules do not lose all birefringence even afterprolonged boiling; swelling power is only about one-fourth and solublesabout one-half that of regular corn starch at 95° C. (Corn and CornImprovement, third edition, Ed. Sprague and Dudley).

High-amylose starches are particularly useful in confectionery such asgummed candies (because they thicken rapidly), in fried snacks (becausethey resist the penetration of cooking oil), and in photographic film(because of their toughness and transparency), as well as in the usesdiscussed above (e.g., textiles, biodegradable packaging materials,adhesives for manufacturing corrugated cardboard, and the like). It hasalso been suggested that the anti-staling properties of bread can beimproved by the use of flour high in amylose. Other uses include thesizing of glass fibers prior to weaving, the preparing of a clear, hotwater dispersible, edible film for packaging food, dyes and othersoluble materials, and coating paper to reduce water and fat absorption.

Nutritional aspects are also important with the high amylose starches,particularly high fiber, high resistance to digestion, low calorie, andcontrol of glycemic response.

Amylopectin is particularly useful in paper-making and adhesives(because its branched chains give it greater binding power), and inready prepared foods, such as in frozen and canned food (because itenhances stability and shelf-life), and fruit pie fillings (where itacts as a thickener). It is useful for the formation of translucentfilms which are readily redissolved, as well as the uses discussedabove.

Other uses of corn include the use of stalks and husks for paper andwall board and the use of cobs for fuel, to make charcoal and for theproduction of fufural.

EXAMPLES

It should be understood that the examples and embodiments describedherein are for illustrative purposes only and that various modificationsor changes in light thereof will be suggested to persons skilled in theart and are to be included within the spirit and purview of thisapplication.

Example 1 Hybrid Comparisons for Agronomic Traits

Comparisons of the agronomic characteristics of PP59601 were made toA5602 which is a commercial amylose hybrid which has similar maturityand adaptation. The comparison is provided in Table 2.

TABLE 2 Hybrid Yield Summary Data for A5602 and PP59601. Comparison datarepresent averages of yield trials conducted over a three year period at4 locations in Indiana. YIELD MST PLTPOP STKLOD TSTWT BU/A 56# PCT PCTPCT LB/BU A5602 151.8 14.4 109.5 2.5 53.1 PP59601 158.7 14.7 101.3 2.051.6 Reps (#) 12 12 12 12 12 Diff 6.9 0.3 8.2 0.5 1.5 (BU/A #56 = numberof bushels per acre of grain yield at 56 lbs per bushel; PCT = percent)

As shown in Table 2, PP59601 has significantly higher agronomic yieldthan A5602. Grain moisture at harvest was comparable indicating that thehybrids have comparable maturities. PP59601 shows a significantadvantage for agronomic yield. PP59601 also had improved stalk qualityover A5602 as indicated by the performance data.

Example 2 Amylose Content

Amylose concentration of the grain was determined by the calorimetricmethod. Amylose selectively absorbs iodine to produce a highly coloredamylose-iodine complex and the intensity of this color is proportionalto the amount of amylose present. The percent transmission is determinedat 610 nanometers using a spectrophotometer. The percent amylose isobtained from a standard curve. This standard curve is prepared from theper cent transmission values obtained with a starch having a known percent amylose content. The calorimetric method used herein is set forthas follows:

Determination of Amylose Content by Colorimetric Analysis

Equipment:

-   -   1. Tecator Cemotec™ sample mill or equivalent    -   2. 4 screw cap glass test tubes with caps, 20×125 mm    -   3. 4 screw cap glass test tubes with cap, 20×150 mm    -   4. 4 solid PTFE (polytetrafluorethylene or teflon) stirring        rods, 8″ in length    -   5. Boiling water bath    -   6. Centrifuge capable of holding 20×125 mm test tubes    -   7. 4 porcelain Büchner funnels, 43 mm plate diameter    -   8. Glass microfiber filters, 4.25 cm diameter, 1-1.2 μm porosity        (Whatman® #1821-042, VWR #28333-141, or equivalent)    -   9. Automated diluter, dual syringe (Hamilton Microlab Series        500® or equivalent)    -   10. Glass syringe for diluter, 10 mL    -   11. Glass syringe for diluter, 500 μL    -   12. Automated flow-injection spectrophotometer, 590 nm        wavelength, such as the Foss Tecator FIAStar™ flow-injection        analyzer system with Tecator 5042 Detector™, Tecator 5012        Analyzer™, Tecator 5027 Sampler™    -   13. Polarimeter, 589 nm wavelength    -   14. Vacuum pump    -   15. Filter flask, 500 mL

Reagents:

Concentrated Calcium Chloride Solution

-   -   3.5 kg of reagent grade calcium chloride dihydrate is dissolved        in purified water, cooled to room temperature, and the specific        gravity adjusted to 1.3 using calcium chloride or purified        water, pH of solution is then carefully adjusted to 2.0 using        reagent grade glacial acetic acid,; solution is filtered through        a medium porosity fritted glass funnel prior to use

Dilute Calcium Chloride Solution

-   -   600 mL of concentrated calcium chloride solution is made up to 2        L with purified water

Stock Iodine Solution

-   -   8.00 g of reagent grade potassium iodide and 4.16 g of reagent        grade iodine is dissolved in approximately 10 mL of purified        water and made up to 1 L with dilute calcium chloride solution;        solution should be stored in an amber bottle

Working Iodine Solution

-   -   25 mL of stock iodine solution made up to 200 mL with dilute        calcium chloride solution\Maize grain sample of known amylose        content to serve as the calibration standard

Procedure:

-   -   1. Finely grind 3-4 g of the calibration standard sample into an        appropriate container using the Cemotec™ sample mill.    -   2. Repeat step 1 for the experimental sample ensuring the mill        is cleaned between the grinding of each sample.    -   3. Weigh 0.2 g, 0.4 g, and 0.6 g of the ground calibration        standard into three separate 20×125 mm test tubes.    -   4. Weigh 0.4 g of the experimental sample into the fourth 20×125        mm test tube.    -   5. Add 8 mL of concentrated calcium chloride solution to each of        the 20×125 mm test tubes.    -   6. Place a PTFE stir rod into each of the 20×125 mm test tubes.        Use the rods to disperse the grain.    -   7. Place the four 20×125 mm test tubes into the boiling water        baths for 30 minutes. Use the stir rods to stir the contents of        the test tubes continuously for the first five minutes. Then        stir the contents for approximately one minute every five        minutes.    -   8. Remove the test tubes from the water bath. Immediately remove        the stir rods without rinsing and allow the samples to cool to        room temperature.    -   9. Add 8 mL of dilute calcium chloride solution to each sample        tube. Cap each tube and shake vigorously.    -   10. Centrifuge the sample tubes at 1,800 RPM for five minutes.    -   11. Carefully place a 20×150 mm test tube into the filter flask        (A sponge can be placed on the bottom of the flask to prevent        breakage of the test tube.)    -   12. Insert the stem of the Büchner funnel into the 20×150 mm        test tube. Place a 1-1.2 μM microfiber filter into the Büchner        funnel. Turn on the vacuum pump.    -   13. Decant the solution from one of the 20×125 mm test tubes off        of the ground grain that was centrifuged to the bottom and onto        the microfiber filter. Allow the sample to filter until all of        the solution has passed into the 20×150 mm test tube and the        filter is dry.    -   14. Cap the 20×150 mm test tube and invert a few times to mix        sample. The filtered solution should be clear and free of        floating particulates at this point. If not, the sample must be        re-filtered.    -   15. Complete steps 11-14 for the experimental sample and for        each one of the calibration standard samples.    -   16. Using an automated dual-syringe diluter, dilute 400 μL of        the filtered sample to 10 mL with dilute calcium chloride        solution.    -   17. Analyze the diluted solutions using a flow-injection        spectrophotometer. The working iodine solution should be used as        the last reagent to be mixed with the injected sample. A        flow-injection pump tube with an inner diameter of 0.38 mm can        be used to deliver the working iodine where as a flow-injection        pump tube with an inner diameter of 0.89 mm can be used to        deliver the sample. If necessary, other reagent bottles filled        with dilute calcium chloride solution can be used with the        flow-injection analyzer to further dilute the sample prior to        mixing with working iodine solution. Purified water should be        used in the rinse station to rinse the flow cell between        analyses.    -   18. Record the peak absorbance value of the iodine treated        solution.    -   19. Using a remaining portion of the filtered solution from step        14, record the optical rotation of each sample.    -   20. Using the results from the three calibration standard        samples, make a plot of Absorbance vs. (Amylose Content×Optical        Rotation.) Determine the slope (m) and y-intercept (b) of this        line.    -   21. Using the values for slope and y-intercept determined in        step 20, and the peak absorbance and optical rotation values for        the experimental sample, the amylose content of the experimental        sample can be determined using the following equation:

${\% \mspace{11mu} {Amylose}} = \frac{{{peak}\mspace{14mu} {absorbance}} - b}{m\mspace{14mu} X\mspace{14mu} {optical}\mspace{14mu} {rotation}}$

TABLE 4 Percent Amylose Content. The following table provides thepercent amylose content of PP59601 compared to commercial hybrids A5602and A5515 for 3 different years at 6 locations in central Indiana.Amylose Content Hybrid Year 1 Year 2 Year 3 Average PP59601 58 56 57 57A5602 57 60 55 57 A5515 59 59 57 58

As shown in Table 4, PP59601 has comparable amylose content comparedcommercial hybrids A5515 and A5602 and falls within the category of aclass 5 amylose hybrid based upon the average amylose content of thehybrid over all trials.

Deposit Information

Applicant has made a deposit on Jun. 2, 2008, of at least 2500 seeds forcorn hybrid PP59601 (as described herein) under the Budapest Treaty withthe American Type Culture Collection (ATCC), Rockville, Md. 20852 USA,ATCC Accession No. PTA-9639. The seeds deposited with ATCC were takenfrom the deposit maintained by National Starch and Chemical Companysince prior to the filing date of this application. This deposit of thecorn hybrid PP59601 will be maintained in the ATCC depository, which isa public depository, for a period of 30 years, or 5 years after the mostrecent request, or for the enforceable life of the patent, whichever islonger, and will be replaced if it becomes non-viable during thatperiod. Additionally, Applicant has satisfied all of the requirements of37 C.F.R. §§1.801-1.809, including providing an indication of theviability of the sample, or will do so prior to the issuance of a patentbased on this application. Applicant imposes no restriction on theavailability of the deposited material from ATCC; however, Applicant hasno authority to waive any restrictions imposed by law on the transfer ofbiological material or its transportation in commerce. Applicant doesnot waive any infringement of rights granted under this patent.

While the invention has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the invention following, in general, theprinciples of the invention and including such departures from thepresent disclosure as come within known or customary practice within theart to which the invention pertains and as may be applied to theessential features hereinbefore set forth and as follows in the scope ofthe appended claims.

Although the foregoing invention has been described in some detail byway of illustration and example for the purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

1. Seed of hybrid maize designated PP59601, a representative sample ofwhich has been deposited under ATCC Accession Number PTA-9639.
 2. Amaize plant, or part thereof, obtainable by growing the seed of claim 1.3. The maize plant, or part thereof, of claim 2, wherein the plant, orpart thereof, have been transformed so that its genetic materialcontains one or more transgenes operably linked to one or moreregulatory elements.
 4. A maize plant having all of the morphologicaland physiological characteristics of the plant of claim
 2. 5. A plantpart having all of the morphological and physiological characteristicsof the plant part of claim
 2. 6. A tissue culture of regenerable cellsproduced from the plant, or part thereof, of claim
 2. 7. A maize plantregenerated from a tissue culture of the plant, or part thereof, ofclaim
 2. 8. An ovule of the plant of claim
 2. 9. Pollen of a plant ofclaim
 2. 10. A method for producing maize seed comprising crossing themaize plant of claim 2 with itself or another maize plant, andharvesting the resultant seed.
 11. The method of claim 10, furthercomprising growing the resultant seed to produce one or more progenymaize plants, breeding from one or more of said progeny maize plants toproduce progeny seed, and harvesting said progeny seed.
 12. The methodof claim 11, further comprising growing said progeny seed, breeding fromthe resultant maize plants to produce seed, and harvesting said seed,over 1, 2, 3, 4, 5, 6 or more generations. 13.-14. (canceled)
 15. A seedwhich when grown produces the plant of claim
 4. 16.-22. (canceled)
 23. Amethod for producing a PP59601-derived maize plant, comprising: a)crossing a hybrid maize PP59601 plant with a second maize plant andharvesting the resultant maize seed, wherein representative seed ofPP59601 has been deposited under ATCC Accession Number PTA-9639; and, b)growing said resultant maize seed to produce a PP59601-derived maizeplant.
 24. A method for developing a maize plant in a plant breedingprogram comprising applying plant breeding techniques to a first maizeplant, or parts thereof, wherein said first maize plant is the maizeplant of claim 4, and wherein application of said techniques results indevelopment of said second maize plant.
 25. The method for developing amaize plant in a maize plant breeding program of claim 24 wherein plantbreeding techniques are selected from the group consisting of pedigreebreeding, recurrent selection, backcrossing, restriction fragment lengthpolymorphism enhanced selection, genetic marker enhanced selection andtransformation.